Analysis Note

ControlIncluded Positive Antigen Control: Catalog # 12-301, non-stimulated A431 cell lysate. Add 2.5 µL of 2-mercaptoethanol/100 µL of lysate and boil for 5 minutes to reduce the preparation. Load 20 µg of reduced lysate per lane for minigels.

Application

Immunoprecipitation:
4 µg of a previous lot immunoprecipitated EGF receptor from 500 µg of A431 RIPA lysate.

Immunocytochemistry:
5 µg/mL of previous lots of the antibody showed positive immunostaining for EGF receptor in A431 cells fixed with 4% paraformaldehyde followed by 100% methanol.

Research CategorySignaling

Anti-EGFR Antibody, clone LA22 is an antibody against EGFR for use in IC, IP & WB.

Research Sub CategoryGrowth Factors & Receptors

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Protein kinases are enzymes that transfer a phosphate group from a phosphate donor onto an acceptor amino acid in a substrate protein. By this basic mechanism, protein kinases mediate most of the signal transduction in eukaryotic cells, regulating cellular metabolism, transcription, cell cycle progression, cytoskeletal rearrangement and cell movement, apoptosis, and differentiation. The protein kinase family is one of the largest families of proteins in eukaryotes, classified in 8 major groups based on sequence comparison of their tyrosine (PTK) or serine/threonine (STK) kinase catalytic domains. Epidermal Growth factor receptor (EGFR) is the prototype member of the type 1 receptor tyrosine kinases. EGFR overexpression in tumors indicates poor prognosis and is observed in tumors of the head and neck, brain, bladder, stomach, breast, lung, endometrium, cervix, vulva, ovary, esophagus, stomach and in squamous cell carcinoma.

Immunogen

A431 membrane proteins

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Linkage

Replaces: 04-337; 04-338

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Protein G Purified

Format: Purified

Purified mouse monoclonal IgG2a in storage buffer (0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl, 0.05% sodium azide). Frozen solution.

Quality

Evaluatd by RIPA lysates from human A431 cells under reducing conditions.

Western Blot Analysis:
0.5-2 µg/mL of this antibody detected EGF receptor in RIPA lysates from human A431 cells under reducing conditions.

Specificity

Recognizes the sequence of Ala351-Asp364 of human EGF receptor, Mr 180kDa, (based on numbering without the signal peptide) (Wu et al., 1989).

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Target description

180 kDa